Effect of Dietary Eicosapentaenoic and Docosahexaenoic Fatty Acid Supplementation during the Last Month of Gestation on Fatty Acid Metabolism and Oxidative Status in Charolais Cows and Calves.

Fatty acids (FAs) are of utmost importance in the peripartal period for the development of the central nervous and immune systems of the newborn. The transport of polyunsaturated fatty acids (PUFAs) through the placenta is considered to be minimal in ruminants. Nevertheless, the cow's FAs are the main source of FAs for the calf during gestation. This research aimed to investigate the influence of low-dose eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) supplementation during late gestation on the FA metabolism of cows and their calves. A total of 20 Charolais cows during the last month of their gestation were included in the feeding trial and were divided into a control group (CON) and an experimental group (EPA + DHA). The latter received a supplement in the amount of 100 g/day (9.1 and 7.8 g/cow/day of EPA and DHA, respectively). Supplementation of low-dose EPA and DHA alters colostrum and milk fatty acid composition through the elevation of n-3 long-chain polyunsaturated fatty acids (LC-PUFAs) without affecting milk fat and protein concentrations and oxidative status. Plasma composition in cows was significantly altered, while the same effect was not detected in calf plasma. No significant change in mRNA expression was detected for the genes fatty acid synthase (FASN) and acetyl-CoA carboxylase alpha (ACACA).

Barcoding (COI) Sea Cucumber Holothuria mammata Distribution Analysis: Adriatic Rare or Common Species?

The overexploitation of the western Pacific Ocean has expanded the sea cucumber fishery into new regions to supply the Asian market. In 2013, sea cucumbers were removed from the Croatian marine protected species list, and commercial fishery took place for a short period (2017-2018) in the Eastern Adriatic Sea. However, holothuroid species are difficult to distinguish. Holothuria mammata is a species that has rarely been reported in this region and strongly resembles the common species Holothuria tubulosa. This is the first study to assess the genetic diversity of sea cucumbers in the Adriatic Sea using genetic barcoding of the mitochondrial gene cytochrome c oxidase subunit 1 (COI). Specimens for barcoding were collected from the northern and central Adriatic, along with a specimen that had been previously identified as H. sp. cf. mammata based on its morphological characteristics. While genetic analyses showed identified this specimen as H. tubulosa, 30% of the collected specimens were genetically identified as H. mammata. These results call into question the historically accepted sea cucumber assemblage in the Adriatic Sea, which regarded H. mammata as a rare species and generally disregarded its presence in large census studies. Such species distribution data are extremely important in developing and monitoring a sustainable fishery.

Detection of Bhanja Bandavirus in Patients with Neuroinvasive Disease of Unknown Etiology in Croatia.

BACKGROUND: Although the Bhanja bandavirus (BHAV) is widely distributed in some European countries, human infections are rarely reported. This study analyzed the prevalence of BHAV antibodies in patients with neuroinvasive diseases of unsolved etiology. METHODS: A total of 254 Croatian patients who developed neurological symptoms during the four consecutive arbovirus transmission seasons (April 2017-October 2021) were tested. Cerebrospinal fluid (CSF) and urine samples were tested using RT-qPCR. In addition, CSF and serum samples were tested using a virus neutralization test. RESULTS: BHAV RNA was not detected in any samples, while neutralizing (NT) antibodies were detected in serum samples of 53/20.8% of patients (95% CI = 16.0-26.3). In two patients, BHAV NT antibodies were detected in the CSF, indicating a recent infection. Both patients were inhabitants of rural areas in continental Croatia, and one reported a tick bite two weeks before symptoms onset. The seropositivity was high in all age groups (15.2-29.1%). The majority of seropositive patients (94.3%) resided at altitudes less than 200 m above sea level. The prevalence rates correlated positively with population density and negatively with certain climate parameters (temperature, number of hot/warm days). CONCLUSIONS: The presented results indicate that BHAV is distributed in Croatia. Further studies are needed to determine the clinical significance of this neglected arbovirus.

Prediction of reproductive performance of ewes based on the early production data, ewe birth rank, dam age, and dam birth rank.

This research aimed to analyze whether ewes' total reproductive performance up to the fourth year of life (RP4) can be predicted based on the data available at an early stage of a ewe's productive life. The RP4 of 133 Romanov ewes was measured in terms of the total number of lambs born per ewe (TNLE) and total birth weight of lambs per ewe (TBLE). Multiple regression was used to analyze whether early reproductive performance indicators (first litter size - FLS, age at first lambing - AFL, first lambing interval - FLI), ewe birth rank, dam age, and dam birth rank can be used as the predictors of RP4. Predicted R 2 and 95 % prediction intervals were used as indicators of the precision of prediction. Average TNLE and TBLE at the end of fourth year of ewe life were 11.84 lambs and 37.96 kg, respectively. FLS and FLI significantly ( P < 0.05 ) influenced TNLE and TBLE, while AFL was not a significant ( P > 0.05 ) variable. Ewes with shorter FLI had significantly ( P < 0.05 ) higher TNLE (10.94 lambs) and TBLE (36.17 kg) than ewes with long FLI (TNLE  =  9.12 lambs and TBLE  =  28.05 kg). R 2 predicted for TNLE and TBLE was 7.54 % and 11.49 %, respectively. The ewe's birth rank and the dam's birth rank significantly ( P < 0.05 ) influenced TNLE and TBLE. Ewes born as singletons and ewes from singleton-born dams had significantly ( P < 0.05 ) lower TNLE and TBLE than ewes born as triplets and ewes from triplet-born dams. R 2 predicted for TNLE was 16.76 %, and 25.69 % for TBLE. FLS and FLI are better predictors of RP4 than AFL. The birth rank of ewe and dam also proved significant predictors of RP4. For both sets of predictors (early reproductive indicators and birth rank data), low values of R 2 predicted indicate that precise prediction of RP4 cannot be made.

A retrospective cohort study of canid herpesvirus-1 seroprevalence and epidemiology in kennel dogs in Croatia.

Canid alphaherpesvirus-1 (CaHV-1) is an endemic pathogen of dogs worldwide. CaHV-1 is often related to abortions, neonatal mortality, and the death of puppies. Since the first description of the virus in 1965, there has been no generally accepted method for diagnosing CaHV-1. Many authors used the virus neutralisation test (VNT) as a 'reference standard' due to its high specificity. Nasal, vaginal, preputial swabs and serum samples for this study were collected in the population of kennel dogs in Croatia. To determine the optimal VNT protocol, three modifications of the VNT were compared. These modifications were: VNT using native serum samples, VNT using thermally inactivated serum samples, and VNT using thermally inactivated serum samples with the addition of the complement. The correlation between the results of the VNT methods was significant (P < 0.001). Of all three modifications of VNT, the one using native serum samples was the one that increased VNT sensitivity. The overall seroprevalence of CaHV-1 was 32.02%. The PCR did not confirm the CaHV-1 presence in collected swabs. According to analysed anamnestic data, significant risk factors for CaHV-1 infection were: kennel size, attendance at the dog shows, hunt trials, kennel disinfection protocols, and mating. The oestrus cycle had no significant effect on seropositivity. The study results indicate that CaHV-1 spreads horizontally between dams living in kennels and in males during mating. Although there was no correlation between seropositivity and a history of reproductive disorders, significantly more stillborn puppies were recorded in seronegative dames (P < 0.01).

Physico-Chemical and Ecotoxicological Evaluation of Marine Sediments Contamination: A Case Study of Rovinj Coastal Area, NE Adriatic Sea, Croatia.

Comprehensive spatial and temporal data on sediment quality in the Adriatic Sea are lacking. Therefore, prior to planned anthropogenic interventions in the local marine environment, such as deepening of the Rovinj harbour, the results of physicochemical and ecotoxicological analyses of five local coastal sediments were compared with regional averages and SQGs of neighbouring countries. Analyses of sediment grain size, content of metals and heavy metals, PAHs and PCBs were performed according to standard protocols. Sediment quality was classified according to French legislation (N1 and N2 level) and sediment guidelines. The phytotoxicity of the eluates was studied by flax seed germination tests. The logistic regression models Pmax and Pavg were used to estimate the probability of toxic effects. Except for the open sea (S5), all other sediments had concentrations slightly higher than the N1 for some metals (Cu, Ni, Hg, Cr) or ΣPAHs, while the Rovinj harbour (S1) reached the N2 value for mercury. The phytotoxicity assay with sediment eluates showed inhibition of germination, root length and root biomass production, with an average phytotoxicity index (PI) ranging from 6.06% to 42.00%. Significant correlations of Pavg and Pmax values with phytotoxicity and other specific parameters were found. In general, according to the applied SQGs, there are no potential ecological impacts on the marine environment at any of the investigated sites, with the exception of site S1.

Kurthia gibsonii, a novel opportunistic pathogen in poultry.

Kurthia gibsonii [H. Kurth, 1883. Ueber Bacterium zopfii, eine neue Bakterienart. Berichte der Deutschen Botanischen Gesellschaft, 1, 97-100] was isolated on 10 unrelated laying hen farms over a period of 15 months. Farmers reported elevated morbidity and mortality rates, and suspected colibacillosis based on the necropsy findings. The most frequently found lesions were perihepatitis, fibrinous peritonitis, salpingitis and oophoritis. Necropsy findings and bacteriological results allowed the diagnosis of colibacillosis. In addition, K. gibsonii was isolated from the ovarian follicles (44.44%), liver (22.22%), peritoneum (16.67%), bone marrow (5.56%), spleen (5.56%), and duodenum (5.56%). On all farms, coinfection with E. coli was detected, while on some farms other common avian pathogens were found as well. In total, 18 K. gibsonii strains were identified and phylogenetically analysed based on the 16S rRNA gene sequences. The results showed some variability of the strains originating from the same farm, although the overall phylogenetic diversity was low, regardless of the geographical location of the farm, age of the flock or date of collection. Embryo lethality assay showed K. gibsonii is not able to cause a primary infection. We conclude that Kurthia gibsonii may play a role as an opportunistic pathogen for poultry. This is the first report of coinfection of Kurthia gibsonii and E. coli in laying hens. RESEARCH HIGHLIGHTSKurthia gibsonii is a novel opportunistic pathogen in poultry.Phylogenetic analysis showed low genetic diversity of Kurthia gibsonii isolates.Embryo lethality assay showed K. gibsonii does not cause primary infections.

Emerging Trends in the Epidemiology of COVID-19: The Croatian 'One Health' Perspective.

During the four pandemic waves, a total of 560,504 cases and 10,178 deaths due to COVID-19 were reported in Croatia. The Alpha variant, dominant from March 2021 (>50% of positive samples), was rapidly replaced by Delta variants (>90%) by August 2021. Several seroprevalence studies were conducted in different populations (general population, children/adolescents, professional athletes, healthcare workers, veterinarians) and in immunocompromised patients (hemodialysis patients, liver/kidney transplant recipients). After the first pandemic wave, seroprevalence rates of neutralizing (NT) antibodies were reported to be 0.2-5.5%. Significantly higher seropositivity was detected during/after the second wave, 2.6-18.7%. Two studies conducted in pet animals (February-June 2020/July-December 2020) reported SARS-CoV-2 NT antibodies in 0.76% of cats and 0.31-14.69% of dogs, respectively. SARS-CoV-2 NT antibodies were not detected in wildlife. Environmental samples taken in the households of COVID-19 patients showed high-touch personal objects as most frequently contaminated (17.3%), followed by surfaces in patients' rooms (14.6%), kitchens (13.3%) and bathrooms (8.3%). SARS-CoV-2 RNA was also detected in 96.8% affluent water samples, while all effluent water samples tested negative. Detection of SARS-CoV-2 in humans, animals and the environment suggests that the 'One Health' approach is critical to controlling COVID-19 and future pandemics.

The Emergence of SARS-CoV-2 within the Dog Population in Croatia: Host Factors and Clinical Outcome.

Over a year into the COVID-19 pandemic, there is growing evidence that SARS-CoV-2 infections among dogs are more common than previously thought. In this study, the prevalence of SARS-CoV-2 antibodies was investigated in two dog populations. The first group was comprised of 1069 dogs admitted to the Veterinary Teaching Hospital for any given reason. The second group included dogs that shared households with confirmed COVID-19 cases in humans. This study group numbered 78 dogs. In COVID-19 infected households, 43.9% tested ELISA positive, and neutralising antibodies were detected in 25.64% of dogs. Those data are comparable with the secondary attack rate in the human population. With 14.69% of dogs in the general population testing ELISA positive, there was a surge of SARS-CoV-2 infections within the dog population amid the second wave of the pandemic. Noticeably seroprevalence of SARS-CoV-2 in the dog and the human population did not differ at the end of the study period. Male sex, breed and age were identified as significant risk factors. This study gives strong evidence that while acute dog infections are mostly asymptomatic, they can pose a significant risk to dog health. Due to the retrospective nature of this study, samples for viral isolation and PCR were unavailable. Still, seropositive dogs had a 1.97 times greater risk for developing central nervous symptoms.